THE MECHANISM OF THE DECOMPOSITION OF CYANAMIDE IN THE SOIL. BY G. A. COWIE. (Rothamsted Experimental Station, Harpenden.) (With Five text-figures.) A PREVIOUS investigation by the writer (i) showed that cyanamide readily breaks down, yielding ammonia in normal clay and sandy soils. The evidence, however, threw no light upon the cause or nature of this change. This question was accordingly reserved for a later investigation. The concensus of the available evidence indicated that the production of ammonia from cyanamide in the soil is due to direct bacterial action. This view was held by Immendorff(2) and Kappen(3), who concluded that in poor soils of low bacterial activities cyanamide is not converted into ammonia but is chemically transformed into dicyanodiamide. L6hnis(4) at first accounted in a similar way for the formation of ammonia from cyanamide in the soil. He (5) assumed later, however, that cyana- mide is normally decomposed by soil colloids into urea or possibly some other substances, and the latter are then converted into ammonia by the soil organisms. He adduced no direct evidence of the production of urea from cyanamide in the soil. Ulpiani (6) regarded the formation of ammonia as primarily due to a purely chemical, not a bacterial, change. He had formerly considered that cyanamide changed into dicyanodiamide in the soil and its value as a fertiliser depended on this change, an opinion also held by Perotti(7). Ulpiani's later work, however, led him to the view that cyanamide breaks down by a purely chemical change to urea which then is con- verted into ammonia. The formation of urea was attributed to the soil colloids. This work was done in culture solutions of various concentra- tions and at various temperatures. Our experiments were made in soil under natural conditions, using amounts of cyanamide comparable with those used in practise. Our results agree with those of Ulpiani. Downloaded from https://www.cambridge.org/core. BBSRC, on 28 Nov 2019 at 14:32:07, subject to the Cambridge Core terms of use, available at https://www.cambridge.org/core/terms. https://doi.org/10.1017/S0021859600006432 164 Decomposition of Cyanamide in the Soil Our experiments have consistently failed to show any appreciable amount of ammonia resulting from the decomposition of cyanamide in sterile soils (heated to 120° C. or 135° C). The addition of the urease of soya-bean, however, produced considerable amounts of ammonia in these soils. This pointed to the presence of urea, which was later con- firmed by the extraction of the soils by alcohol and identification of urea in the extract by the urea-nitrate test. Further experiments more- over demonstrated that urea actually remains stable in soils heated to 120° C. The addition of cyanamide to sterile soils thus leads to an accumulation of urea, which persists as such in consequence of the suppression of the necessary urea decomposing organisms. In a similar way the addition of cyanamide to soils heated to 100° C. does not lead to an immediate production of ammonia. It forthwith yields urea, however, which then decomposes into ammonia after the. recovery of the appropriate organisms. The evidence shows, on the other hand, a rapid and progressive production of ammonia arising from the decomposition of cyanamide in unheated normal clay and sandy soils. Careful examination of these soils, however, revealed in the initial periods the presence of appreciable amounts of urea. This indicates that urea produced by the decomposition of cyanamide also accumulated to some extent under normal conditions. The cumulative evidence thus leads to the conclusion that cyanamide in the soil is normally converted by a purely chemical process into urea and this change is not dependent on the activity of micro-organisms. The urea is then broken down to ammonia by a change which, as the curves indicate, is produced by soil organisms. Cyanamide appears to behave in this way in both clay and sandy soils, but the decomposition seems to be more rapid in the former than in the latter. The experiments have conclusively shown that cyanamide does not decompose into urea in ordinary impure quartz sand; whatever the decomposing agent may be it is not present in pure sand. Cyanamide does not appear to break down in the manner above indicated in peat and fen soils; in these it gives rise to a relatively small production of urea under normal conditions. The investigation has not revealed the exact nature of the decom- posing agent in the soil. It is interesting to note, however, that a sample of Thanet sand taken from a boring through the London Clay near Chelmsford was found even after ignition to be active in decomposing cyanamide into urea. This particular sand (8) has been shown to contain a constituent resembling a zeolite in being reactive and possessing the Downloaded from https://www.cambridge.org/core. BBSRC, on 28 Nov 2019 at 14:32:07, subject to the Cambridge Core terms of use, available at https://www.cambridge.org/core/terms. https://doi.org/10.1017/S0021859600006432 G. A. COWIE 165 property of softening hard water by the substitution of sodium salts and possibly potassium for those of calcium and magnesium. In following up this clue it was found that the addition of a definite zeolite prehnite to ordinary inert sand produces a mixture capable of converting cyana- mide into urea. EXPERIMENTAL. The cyanainide was used in the form of fresh nitrolim in which the calcium cyanamide had undergone practically no change. The bulk of the soil to be used was first thoroughly mixed together and its moisture content raised, where necessary, to 12 to 15 per cent, according to its water capacity. The soil was next passed through a 3 mm. sieve, weighed out into lots of 200 grams which were then transferred to wide-mouthed bottles of 10 oz. capacity, a fresh bottle being taken for each deter- mination at the end of the various periods. The application of the cyanamide to the soils was made in the following manner. In the case of unheated soils the weighed quantity of cyanamide was sprinkled on to the 200 grams of soil previously spread out on a sheet of paper; the whole was then mixed and replaced in the bottles, which were plugged with cotton-wool. In the case of the heated soils this procedure might have led to reinfection, and therefore the bottles containing soil, etc. were first plugged with cotton-wool and then heated for the proper period at the requisite temperature. After cooling, the cyanamide was carefully and rapidly introduced into the bottles from small glass tubes, in which the cyanamide itself had been heated for the same time at the same tem- perature in the air-oven. Tests showed that heating under these con- ditions produced no chemical change in the cyanamide. After careful replugging to avoid reinfection the bottles were vigorously shaken for some considerable time to ensure an adequate mixing of the cyanamide with the soils. In no case did heating cause a loss of more than 2 per cent, moisture in the soils. All bottles were then kept in a dark cellar at the ordinary laboratory temperature. In order to inhibit the nitrification of ammonia produced from cyanamide, a small amount of dicyanodiamide, equivalent to thirty parts N per million dry soil, was added with the cyanamide. A previous investigation showed conclusively that dicyanodiamide does not give rise to ammonia or affect appreciably the production of ammonia from cyanamide. Downloaded from https://www.cambridge.org/core. BBSRC, on 28 Nov 2019 at 14:32:07, subject to the Cambridge Core terms of use, available at https://www.cambridge.org/core/terms. https://doi.org/10.1017/S0021859600006432 166 Decomposition of Cyanamide in the Soil THE STAGES IN THE PRODUCTION OF AMMONIA FROM CYANAMIDE IN SOILS. Experiments were made to determine the relative rates of ammonia production from cyanamide in untreated soils and the same soils after heating for one half hour in the autoclave at 120° C. Both the heavy Rothamsted and the light Woburn soils were used. The results (plotted in Fig. 1 and given in Table I) show a rapid and progressive production of ammonia from cyanamide in the unsterilised soils, but they afford little evidence of ammonia resulting from the decomposition of cyana- mide in the sterilised soils. 100r Rothamsted (Heated) 0 13 5 10 20days Fig. 1. Showing relative rates of ammonia production from cyanamide in unheated soils and soils heated to 120° C. Although the sterilised soils showed little trace of ammonia pro- duction from cyanamide, there was the possibility that the decomposition had stopped at some intermediate stage. On chemical grounds the most probable stage seemed to be the formation of urea. Examination for this substance was therefore made in soil which had been stored for 20 days or more, the urease of the soya-bean being used as the testing Downloaded from https://www.cambridge.org/core. BBSRC, on 28 Nov 2019 at 14:32:07, subject to the Cambridge Core terms of use, available at https://www.cambridge.org/core/terms. https://doi.org/10.1017/S0021859600006432 G. A. COWIE 167 agent. Takeuchi(9) and also Armstrong and Horton(iO) have shown this enzyme is quite specific in its action, decomposing urea only, and nothing else so far as is known. The soils were accordingly treated with Table I. Ammonia produced from Cyanamide in Soils. N present as NH3 per million dry soil Rothamsted Woburn Control at start 4-0 Control at start 2-6 After After After After After After After After After After 1 3 5 10 20 1 3 5 10 20 Treatment day days days days days day days days days days Control 4-0 3-4 4-0 6-7 6-7 2-6 1-9 2-6 5-2 5-2 + Cyanamide 44-5 82-3 90-3 971 97-1 22-0 55-5 87-8 98-2 100 Heated soil 9-3 19-9 23-9 25-2 25-2 7-7 7-7 191 17-9 21-7 (120° C.) + Cyanamide 14-6 14-6 29-2 31-9 30-5 12-8 140 230 25-5 26-8 Cyanamide N added = 100 parts per million dry soil.